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. 1982 Aug 11;10(15):4467–4482. doi: 10.1093/nar/10.15.4467

Chemical synthesis and cloning of a gene for human beta-urogastrone.

J Smith, E Cook, I Fotheringham, S Pheby, R Derbyshire, M A Eaton, M Doel, D M Lilley, J F Pardon, T Patel, H Lewis, L D Bell
PMCID: PMC321104  PMID: 6290982

Abstract

A DNA duplex coding for the 53 amino acids of human beta-urogastrone has been synthesised. Computer assisted design of the gene included restriction endonuclease sites for plasmid insertion, a termination codon and two triplets coding for lysine at the 5'-end of the structural gene. The synthesis involved preparation of 23 oligodeoxyribonucleotides by phosphotriester procedures coupled to rapid HPLC techniques. The gene was constructed in two halves by enzymatic ligation of the oligonucleotides and cloned into a specially constructed chimeric plasmid vector. Escherichia coli K12 MRC8 was transformed by the plasmid and clones containing the full gene sequence were isolated and characterised.

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Selected References

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